issn no.: 2454- 2024 (online) international journal of ... fungi and... · widely [57]. dead sea,...

ISSN No.: 2454- 2024 (online)

International Journal of Technical Research & Science

pg. 345

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Paper Id: IJTRS-V1-I10-011 Volume 1 Issue 10, January 2017

@2016, IJTRS All Right Reserved

EXTREMOPHILIC FUNGI AND CHEMICAL

ANALYSIS OF HYPERSALINE, ALKALINE

LAKES OF WADI-EL-NATRUN, EGYPT Ismail, M. A.

1,2, Moubasher, A. H.

1,2,3, Mohamed A. Ramadan

1 and Al-Bedak, O. A.

2

1Botany and Microbiology Department, Faculty of Science, Assiut University, 71515, Assiut, Egypt 2Assiut University Mycological Centre (AUMC), Assiut University, 71515, Assiut, Egypt

3Corressponding author: [email protected]

Abstract- Mesophiles, halophiles and thermophiles fungi were isolated from water, mud and newly reclaimed

soil of the biggest eight hypersaline, alkaline lakes of Wadi-El-Natrun, Egypt. Morphological techniques were

used to identify the isolated fungi and in many cases molecular technique was used to confirm the identification.

Mesophiles showed the highest number of taxa from newly reclaimed soil, followed by mud and water.

Aspergillus was almost the most common genus recovered on the three isolation media contributing regularly

the highest CFUs. Acremonium was the most dominant genus recovered from water on Czapek's + 10 % NaCl

medium. Penicillium was the second most dominant behind Aspergillus in the mud and soil and it was

completely absent at 45° C. A distinctive isolate was recovered from a water sample representing a new species

of Ramophialophora based on its morphology and rDNA analysis. A 281 strains of mesophiles and halophiles

were screened for their ability to grow on Czapek’s broth medium supplemented with different NaCl

concentrations (5, 10, 15, 20, 25 and 30 %) and all could grow up to 15 % NaCl (weakly halotolerant), while

248 isolates up to 20 % NaCl (moderately halotolerant), 32 up to 25 % NaCl (highly halotolerant) and 12 could

grow up to 30 % NaCl (extremely halotolerant). These 12 strains could further survive 30 % salt concentration

for up to18 months.

Key words: Alkaline lakes, Extremophiles, Halophiles, Hypersaline, Thermophiles, Wadi-El-Natrun.

1. INTRODUCTION

Hypersaline lakes are characterized by extreme concentrations of NaCl, often high concentrations of other ions,

and in some cases extremes in pH. After evaporation, soda soils are formed with high pH values (> 8) which

represent an example of extreme habitats. Soda soils usually develop in arid and semi-arid lands worldwide, and

may be varied in salt concentrations from low to saturation [28]. The driving force for the soda accumulation is

the loss of Ca+2

trapped by CO3-2

ion, leaving Na+ as the dominant cation [36]. Soda soils are formed as a result

of carbonate accumulation under poor Ca+2

and Mg+2

conditions [28, 36].

Hypersaline waters (lakes) can be divided into thalassohaline and athalassohaline, the first have a marine origin

as they have a composition similar to that of sea water and the second, whose composition reflects the

composition of the surrounding geology, topography and environmental conditions and its composition is varied

widely [57]. Dead Sea, Great Salt Lake, some cold hypersaline lakes in Antarctica or alkaline lakes such as Lake

Magadi or the lakes of Wadi-El-Natrun are typical examples of athalassohaline waters [27, 57].

Fungi are globally distributed over a wide range of ecological extreme habitats, and to conquer them they must

be able to rapidly germinate and become established to produce the necessary extracellular enzymes in the

encirclement environment [59]. The study of such these extremes are very important for the egression of new

species and evolution of various ecological relations among organisms which compensate certain environmental

extremes. New metabolites and metabolic pathways of organisms from such rigorous habitats can be expected

[52].

These extreme habitats are occupied by highly specialized organisms which termed extremophiles. Halophiles

are considered an important organisms that existed in hypersaline habitats and they can be divided on the basis

of their optimal growth as 1) non-halophiles are those that grow best in media containing less than 1.16 % NaCl,

2) slightly halophiles grow best in media with 1.16-2.9 %, 3) moderately halophiles grow best with 2.9-14.5 %

and 4) extremely halophiles show optimal growth in media containing 14.5-30.16 % [39]. Another classification

proposed was based on the halophilic abilities of the tested fungi which categorized them as 1) halophilic, which

could grow on 5-25 % NaCl, 2) highly halotolerant, which could grow on 0-20 %, 3) moderately halotolerant,

which could grow on 0-15 % and 4) weakly halotolerant, which could grow on 0-10 % [47].

1.1 Objectives of the Study

To study the chemical analysis of the water, mud and newly reclaimed soil around the hypersaline,

alkaline lakes of Wadi-El-Natrun.

To study the biodiversity of the mesophilic, halophilic/halotolerant and thermophilic/thermotolerant

fungi that inhabit these habitats.

To assess the mesophilic and halotolerant/halophilic fungal isolates for their ability to survive different

levels of NaCl and different storage periods.



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2. MATERIALS AND METHODS

2.1 Site Description

Wadi-El-Natrun (“Natrun” is derived from the Latin word “natrium” for the element sodium) is a part of the

Western Desert adjacent to the Nile Delta and belongs to Behira Governorate, Egypt. It is a narrow depression

located approximately 90 km south of Alexandria and 110 km northwest of Cairo between Lat. 30º17' and 30º38'

N and between Long. 30º2' and 30º30' E [65]. It is narrow at both ends (2.6 km in the north and 1.24 km in the

south) and wider in the middle (8 km) and lies 23 m below sea level and 38 m below the water level of Rosetta

branch of the Nile [7]. It is characterized by a series of twenty small disconnected shallow lakes in its bottom

aligned with its general axis in the northwesterly direction for a distance of about 30 km, most of them no deeper

than half a meter and they have become hypersaline by evaporative concentration [32]. These lakes are fed by

two sources: the springs in the bottom (e.g. in Lake Hamra) and the underground seepages from the Nile River

[32]. Groundwater (the source of the salts in the lakes) derived from the Nile delta infiltrates the Wadi in small

trickles due to the presence of a hydrostatic connection between the Nile (Rosetta branch) and the Wadi [32, 60].

2.2 Samples Collection

40 water, 40 mud (5 samples/each lake) and 33 newly reclaimed soil samples were collected during February

2012 from the 8 main biggest lakes in the Wadi, namely Fasida, Umm-Rishah, Rosetta, Hamra, Zugm, Beida,

Khadra and Gaar.

2.3 Sample Preparation for Chemical Analysis

As soon as samples were collected, they were transferred to the laboratory. Moisture content was determined for

all the mud and soil samples investigated before sample preparation for chemical and mycological analysis. The

mud and soil samples were spread out on separate polyethylene sheets and left to air dry for 3 days. The dried

samples were ground by means of wooden mortar and pestle to reduce particle size to pass through 2-mm sieve,

then were thoroughly mixed and put in plastic bags and preserved in a refrigerator at 4°C till chemical and

mycological analysis.

2.4 Chemical Analysis

pH, organic matter content [56], total dissolved solids (TDS) [67], sodium and potassium [15], calcium and

magnesium [14], carbonates, bicarbonates and chlorides [66] were determined for all samples investigated.

2.5 Media Used for Isolation of Fungi

1 % glucose-Czapek’s agar medium of the following composition (g/l): glucose, 10; Na2NO3, 2;

K2HPO4, 1; KCl, 0.5; MgSO4.7H2O, 0.5; FeSO4, 0.01; ZnSO4, 0.01; CuSO4, 0.005, agar, 15 and final

pH 7.3.

1 % glucose–Czapek’s agar of the above composition + 10 % NaCl.

Yeast-Starch agar (YpSs) [24] of the following composition (g/l) of: Soluble starch, 15; powdered

yeast extract, 4.0; K2HPO4, 1.0; MgSO4.7H2O, 0.5; agar, 15; water (1/4 tap and 3/4 distilled), up to

1000 ml. and final pH 7.0. To each of these media, Rose Bengal at 0.05 g/l and chloramphenicol at 250

mg/l [10, 63] were added to suppress the growth of bacteria and to restrict the fungal colonies which

facilitate the isolation of slow-growing fungi.

2.6 Isolation of Fungi from the Water Samples

Pour plate technique was used for isolation of fungi from the water samples. Three ml of each water sample

were pipetted into each 9.0 cm Petri dish (5 dishes/sample). Three isolation media and two incubation

temperatures were used for isolation of mesophiles, halophiles and thermophiles. The inoculated plates were

incubated for 7-15 days at 25ºC for mesophiles and halophiles and at 45ºC for thermophiles fungi. Counts of

CFUs of each fungus were calculated per 600 ml water in all samples.

2.7 Isolation of Fungi from the Mud and Soil Samples

Dilution plate method [34] was used for isolation of fungi from mud and soil samples. The inoculated plates

were incubated for 7-15 days. Counts of CFUs of each fungus were calculated per 1 gm oven-dry mud or soil in

every sample. Five plates were used for each treatment.

2.8 Halophilic Activity



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This test was performed using Czapek’s solution supplemented with NaCl concentrations of 0, 5, 10, 15, 20, 25

and 30 % [47]. 281 fungal strains (144 from saline water, 37 from mud and 100 from soil) were tested. 15 ml of

Czapek’s solution was transferred into 32-ml screw-capped tubes, autoclaved at 121° C for 20 min and

inoculated with spore mass of 7-day-old culture of the tested strains and incubated at 25° C for 15-60 days. The

visual growth was recorded for positive strains. The growth of the highest halophiles strains that grow at 30 %

NaCl was tested on Cz agar after different storage periods of 3, 6, 9, 12, 15 and 18 months.

2.9 Phenotypic Identification of Fungi

The identification of fungal genera and species was based on macroscopic and microscopic features

following the keys and descriptions of Aspergillus species [55], Penicillium and its teleomorphs [54],

Dematiaceous Hyphomycetes [21, 22], Fusarium species [11, 40], fungi in general [19, 42] and thermophilic

and thermotolerant fungi [16, 58].

2.10 Molecular Identification of the Isolated Fungi

The fungus was grown on Czapek’s yeast extract agar (CYA) plates and incubated at 25° C for 7 days. A small

amount of the fungal growth was scraped and suspended in 200 µl of distilled water and boiled at 100° C for 15

minutes, and sent to SolGent Company, Daejeon, South Korea, for PCR and rDNA sequencing.

Fungal DNA was extracted and isolated using SolGent purification bead in SolGent Company (SolGent,

Daejeon, South Korea). Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA were amplified

using the universal primers ITS 1 (5' - TCC GTA GGT GAA CCT GCG G - 3'), and ITS 4 (5'- TCC TCC GCT

TAT TGA TAT GC -3'). Then amplification was performed using the polymerase chain reaction (PCR) (ABI,

9700). The PCR reaction mixtures were prepared using Solgent EF-Taq as follows: 10X EF-Taq buffer 2.5 µl,

10 mM dNTPs (T) 0.5 µl, primer (F-10p) 1.0 µl, primer (R-10p) 1.0 µl, EF-Taq (2.5U) 0.25µl, template1.0 µl,

distilled water to 25 µl. Then the amplification was carried out using the following PCR reaction conditions: one

round of amplification consisting of denaturation at 95 °C for 15 min followed by 30 cycles of denaturation at

95 °C for 20 sec, annealing at 50 °C for 40 sec and extension at 72 °C for 1 min, with a final extension step of

72 °C for 5 min.

The PCR products were then purified with the SolGent PCR Purification Kit-Ultra prior to sequencing. Then the

purified PCR products were reconfirmed (using size marker) by electrophoreses of the PCR products on 1%

agarose gel. Then these bands were eluted and sequenced. Each sample was sequenced in the sense and

antisense direction.

Contigs were created from the sequence data using CLCBio Main Workbench program. The sequence obtained

from each isolate was further analyzed using BLAST from the National Center for Biotechnology Information

(NCBI) website. Sequences obtained with those retrieved from GenBank database were subjected to Clustal W

analysis using Meg Align (DNA Star) software version 5.05 for the phylogenetic analysis. Sequence data were

deposited in GenBank and accession numbers are given for them.

3. RESULTS AND DISCUSSION

3.1 Chemical Analyses

3.1.1 Water samples

In the present study, the water samples showed alkaline nature of pH ranging from 8.5 to 9.54 (Table 3.1). In

agreement with the current results nearly the same range of pH (8.51- 9.45) was registered in the water of Wadi-

El-Natrun lakes [9, 26, 48, 65], (9.4 - 9.8) in the water of Mono Lake, California [64], (7.9 - 9.1) in the water of

Sutton Salt Lake, east Otago, New Zealand [17], and (8.66 - 9.27) in the water of the Chaplin Lakes [12]. In the

current study, the water samples contained TDS ranging from 1.92 % in Lake Khadra to 26.3 % in Lake Umm-

Rishah (Table 3.1) and these TDS range was disagreed with the higher levels of TSS previously recorded in the

water of Wadi-El-Natrun lakes during 2006/2007 [9, 26, 65]. These differences may be due to the difference in

the season of sampling. In agreement with these results almost similar TDS levels (9.0 %) in Mono Lake in

California and (8.8 %) in Big Soda Lake in Nevada were obtained [53]. Also values of TDS ranging from 2.487

% in Lake Big Quill to 25.18 % in Lake Ingebright were recorded [12].

In the current investigation, the water samples registered sodium cations and chloride anions of (1.13 - 17.64

g/l) and (0.72 - 13.52 g/l) respectively. In contention with these results, higher levels were reported in the water

of Wadi-El-Natrun lakes ranged from 17 - 44 g/l of sodium and 10.5 - 23 g/l of chloride [9, 48]. Also higher

contents of sodium and chloride of (17.6 g/l and 29.5 g/l respectively) in Mono Lake in California and (28.0 g/l

and 27.0 g/l respectively) in the Big Soda Lake in Nevada were recorded [53]. However much higher levels of

sodium (78.41 g/l) and chloride (47.54 g/l) were reported in Lake Katwe in Uganda [38] and (30.8 g/l and 49.1

respectively) in Sevier Lake Utah [30].

Carbonates showed its peak (38.1 g/l) in Lake Umm-Rishah while the highest value of bicarbonates (29.2 g/l)

was reported in Lake Fasida (Table 3.1). This high level of carbonates and bicarbonates may explain the high



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alkalinity of Lake Fasida and Lake Umm-Rishah. These values are much higher than those reported in the water

of Wadi-El-Natrun lakes during 2006/2007 which registered lower values of carbonates (0.2 - 1.3 g/l) and

bicarbonates (0.11- 2.0 g/l) [9, 48]. Also, a higher sum of carbonates and bicarbonates of 30.1 g/l in Mono Lake

and 24.0 g/l in the Big Soda Lake was recorded [53].

Other cations: K+

(0.0186-0.45 g/l), Ca+2

(0.03 - 0.64 g/l) and Mg+2

(0.01- 0.09 g/l) were found at very low

levels compared to those reported in the Great Salt Lake, Dead Sea, Sevier Lake Utah and ocean water [30]. In

agreement with the current results, similar concentrations of calcium (0.15 g/l), magnesium (0.3 g/l) and

potassium (0.6 g/l) in the water of Wadi-El-Natrun lakes were previously found [50]. Also lower levels of

potassium (0.1- 0.3 g/l), calcium (0.01- 0.5 g/l) and magnesium (0.02 - 0.3 g/l) in the water of Wadi-El-Natrun

lakes were reported during 2006/2007 [9, 48].

3.1.2 Mud samples

In the present study, pH of the mud samples showed its peak (10.24) in Lake Fasida while the least (8.78) was

recorded in Lake Rosetta (Table 3.2). In this respect, similar results of pH of the mud samples of Wadi-El-

Natrun lakes ranging from 9.0 to 9.4 [9, 26], the mud samples of Mono Lake ranging from 8.6 - 9.45 [64] and

Playa Lake sediments ranging from 8.5 - 9.5 [61] were previously recorded.

In the current study organic matter content ranged from 0.45 % in Lake Fasida to 5.94 % in Lake Zugm, and this

was higher than those recorded from the mud samples of Wadi-El-Natrun lakes during 2006-2007 [9, 26]. TDS

values ranged from 0.66 % in Lake Khadra to 20.36 % in Lake Rosetta mud, however higher TSS values ranged

from 15.8 - 38.3 % in the mud samples of Wadi-El-Natrun lakes during 2006-2007 were registered [9, 26].

Sodium cation (g/kg) in the mud registered a range of 76.5 in Lake Khadra to 281.11 in Lake Beida which is

much higher compared to those previously recorded during 2006/2007 in the mud samples of Wadi-El-Natrun

lakes [9, 26] and those reported in the mud samples of Mono Lake, California [64].

The current study showed that Lake Hamra registered the peak of potassium cations (7.1 g/kg) while the trough

(0.97) was recorded in Lake Khadra. However, relatively lower levels of 0.2 g/kg in Lake Fasida and 2.3 g/kg in

Lake Zugm were previously reported in the mud of Wadi-El-Natrun lakes during 2006/2007 [9, 26]. Nearly

similar results were obtained from the sediments of Lake Abert, Oregon, USA which contained 1.7 - 7.7 g/kg

potassium [35] while the mud samples of Mono Lake registered much lower potassium content ranging from 0-

0.265 g/kg [64].

Low calcium amounts of 0.18 - 2.3 g/kg and magnesium of 0.05 - 0.17 g/kg were registered in the mud samples

(Table 3.2) and this is concordant with those previously obtained from the mud of Wadi-El-Natrun lakes during

2006/2007 where low calcium (0.01 - 0.1 g/kg) and magnesium (0.02 - 0.2) content was recorded [9, 26].

Higher values of magnesium content ranging from 16.1-41.5 g/kg have been reported from sediments of Lake

Abert [35]. Much higher calcium content of 18.95-38 g/kg and magnesium of 1.5 - 1.75 g/kg have been reported

in the mud samples of Mono Lake, California [64].

In the present study, carbonates, bicarbonates and chlorides yielded their peaks (22.98, 16.06 and 90.9 g/kg

respectively) in the mud of Lake Rosetta which registered the highest TDS while their trough (0.12, 1.67 and 2.5

g/kg respectively) were recorded in Khadra which recorded the least TDS level. These current results are in

disagreement with the previously reported range of carbonates (0.02 - 0.45 g/kg), bicarbonates (0.18 - 0.76) and

chlorides (5.7 - 16.4) in the mud of Wadi-El-Natrun lakes during 2006/2007 [9, 26].

3.1.3 Soil samples

In the present study, the soil samples collected from around the lakes exhibited slightly alkaline pH values

ranging from 7.83 to 8.12 which harmonize with the results previously obtained from soil samples around lakes

of Wadi-El-Natrun [9, 20, 26, 49] and soil around the Mono Lake, California [64]. The soils around lakes of

Wadi-El-Natrun are newly reclaimed and depend on humus and different types of fertilizers which may explain

its high range of organic matter (1.5 - 2.49 %) compared with that previously recorded from different types of

soil in Egypt [47] and Libya [23].

In the current investigation, moisture content of the soil samples ranged from 6.9 % in Lake Khadra to 12.6 % in

Lake Fasida. Other parameters (TDS, Na+, K

+, CO3

-2, HCO3

-, Ca

+2, Mg

+2, Cl

-) were reported in lower levels in

the soil than those registered in the water and mud (Table 3.3) and this may explain the highest number of

propagules in the soil than in the water and mud, since stresses from pH, TDS, Na+ and other cations and anions

are much lesser. A slightly higher range of TSS (0.5 - 6.2 %) during 2006/2007 was reported [9, 26]. However,

soils of Mono Lake area revealed much higher content of chlorides (53.25 - 61.75 g/kg), calcium (50.85 - 53.05)

and magnesium (2.45 - 2.95) while potassium was at zero level and sodium ranged from 0.0 to 1.85 [64].

3.2 Biodiversity of Fungi



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Three media and two incubation temperatures were employed. 44 genera, 129 species and 4 varieties were

recorded from the water, mud and newly-reclaimed soil samples on the three media at 25º and 45º C. As

expected, the CFUs were consistently higher on Cz than on Cz + 10 % NaCl at 25º C and YpSs at 45° C. In the

hypersaline waters of Wadi-El-Natrun lakes the mean density of fungi was 76.66 cells/l on Cz + 10 % NaCl at

25° C, 78.33 cell/l on YpSs at 45° C and 831.66 cells/l on Cz at 25° C. Fungal counts of about 10 cells/l were

typical for ocean water [62]. On Cz at 25º C, the mud was the richest source in fungal genera (28) while the

water was the richest in fungal species (61 + 1 variety). On Cz + 10 % NaCl, mud gave the highest number of

genera (8) while soil contributed the highest number of species (27 + 1 variety). Soil was also the richest in

fungal genera (14) and species (19 + 3 varieties) on YpSs at 45º C.

Identification of fungi was based mainly on the macroscopic and microscopic characteristics, but in many

suspected cases molecular methods (ITS sequence analysis of the rDNA) were used to confirm their

identification (Table 3.4).

3.2.1 Mesophiles

A total of 105 species and 1 variety appertaining to 35 genera were isolated from the three habitats. Aspergillus

contributed a total of 23 species and one variety from the water, mud and soil. The biodiversity of Aspergillus

species was more pronounced in water (18 species + 1 variety) than in mud and soil (13 species + 1 variety

each). A. fumigatus was the most dominant species from water followed by A. sulphureus which contributed

7.21 % and 6.61 % of total fungi respectively (Table 3.5). A. terreus was the most dominant species from soil

(26.15 %) and mud (12.8 % of total fungi). A. flavipes came next to A. terreus in its dominance in mud (11.2 %

of total fungi) and A. flavus came next in soil (6.67 % of total fungi). Penicillium was the runner up of

Aspergillus, represented by 20 species recovered from the three habitats. Similarly, Penicillium from the water

yielded more species (12) than the mud (10) and soil (5) (Table 3.5).

The current results are in agreement with those previously recorded from water samples collected from the 8

lakes of Wadi-El-Natrun during 2006/2007 where 15 species related to 7 genera were reported and it was found

that Aspergillus was the most dominant genus followed by Acremonium and Penicillium [48]. Also 81 species

and 2 varieties from the alkaline soil around Wadi-El-Natrun lakes were recorded and it was found that

Aspergillus, Fusarium, Myrothecium, Stachybotrys, Penicillium and Emericella were the most common genera

[49]. Also, these results are in accordance with those recorded from alkaline soil (pH 8.1-8.6) around Basrah,

Iraq where Aspergillus and Penicillium were the most common fungal genera isolated [33].

Some Aspergillus species have been recorded from the water of solar salterns of Cabo Rojo in Puerto Rico,

namely A. caespitosus, A. candidus, A. flavipes, A. flavus, A. melleus, A. nidulans, A. ochraceus, A.

penicillioides and A. unguis [13]. Penicillium citrinum, P. chrysogenum, P. oxalicum and P. variabile were also

isolated from water of solar saltern of Cabo Rojo in Puerto Rico, while A. japonicas, Chaetomium globosum,

Cladosporium cladosporioides and P. variabile were recorded from sediments [13]. It was found that A. flavus,

A. janus, A. japonicas, A. niger, A. sydowii and A. terreus were the most common species from alkaline soil

samples collected from Casa Caiada and Bairro Novo Beaches, Brazil [25]. Aspergillus and Penicillium were

also isolated in high frequencies from salt marsh soils in Egypt where A. flavus, A. fumigatus, A. niger and A.

ochraceus were the most common species [6].

Some fungal species were recorded for the first time from hypersaline alkaline water in Egypt; these were

Acremonium alternatum, A. zonatum, Alternaria chlamydosporigena, Aspergillus insulicola, A.

roseoglobulosus, Chordomyces antarcticum, Penicillium dendriticum, P. melinii, P. sublateritium,

Phaeoacremonium sp., Plectosphaerella oligotrophica and Stilbella fimetaria. A distinctive isolate was also

recovered from the water of Lake Fasida probably representing a new species of Ramophialophora based on

morphology and rDNA analysis (Tables 3.4, 3.5).

3.2.2 Halophiles

In the present study, 38 species and one variety related to 11 genera were isolated on Cz + 10 % NaCl at 25° C

from the three habitats contributing a total of 3670 CFUs per 1 g or 1 ml in all samples. The soil was the richest

in the number of species (27 species and 1 variety) followed by mud (17 species) while the water registered the

least number (3 spp.). Aspergillus was the most frequent genus obtained from the soil (93.9 % of the samples)

followed by mud (32.5 %) contributing 81.0 % and 85.46 % of total fungi respectively and was missing in water

(Table 3.6). A. flavipes was the most dominant species recovered from mud samples contributing 36.8 % of

total fungi and A. flavus was the most dominant from the soil comprising 37.2 % of total fungi and this is in

agreement with the previously obtained results from soil of Wadi-El-Natrun lakes [49]. 14 species of

Penicillium were isolated from desert soils of Saudi Arabia on 5 % NaCl agar medium [2]. It was found that P.

chrysogenum and P. citrinum were among the common species in soils from the Red Sea shore and Wadi Bir-

El-Ain on 5-10 % NaCl agar [8, 46]. Acremonium was the most dominant fungus from water samples exhibiting

93.48 % of total fungi (Table 3.6).



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Previous studies revealed that at low water content the active mycobiota were dominated by species of

Aspergillus and Penicillium, which were the most common taxa [18] as observed in studies of Mono Lake,

California [64], Arctic [29] and Cabo Rojo in Puerto Rico [13]. Only 2 species of Scopulariopsis (S. halophilica

and S. brumptii) and Acremonium hyalinulum were previously recorded from water samples during one season

of study of the water of Wadi-El-Natrun lakes [48]. However, 41 species and one variety related to 16 genera

were isolated from the soil samples collected from around lakes of Wadi-El-Natrun during 2006/2007 [49] and

it was concluded that the majority of fungi of Wadi-El-Natrun are osmophiles and halophiles and they are able

to accommodate themselves to exist under conditions of water stress but a lower number can tolerate water

stress with the toxic effect of excessive concentrations of Na+ and Cl

- ions.

3.2.3 Thermophiles

Twenty-five species and 3 varieties belonging to 14 genera were isolated from water, mud and soil samples on

YpSs at 45°. Soil was the richest source yielding 14 genera, 19 species and 3 varieties. Aspergillus was the most

dominant genus in the mud and soil represented by 3 species each contributing 30.2 % and 17.76 % of total

fungi respectively (Table 3.7). In this respect, almost similar results were obtained from the soil, mud and salt-

crusts of Wadi-El-Natrun lakes [9] where it was found that Aspergillus (6 species + 1 variety) and Emericella (5

species) were the most common genera at 45º C. Thermomyces represented by 2 species; T. ibadanensis which

was isolated from the mud and soil comprising 4.14 % and 4.5 % of total fungi respectively and T. lanuginosus

which was recovered from the water, mud and soil comprising 14.9 %, 11.83 and 7.65 % of total fungi

respectively.

Previous studies of thermophilic and thermotolerant fungi in Egypt revealed that A. fumigatus, Chaetomium

thermophile, Emericella nidulans, Malbranchea cinnamomea and Talaromyces thermophilus, were recorded

from cultivated and desert soils in Egypt [31] and from desert soils in Saudi Arabia [3] and Myriococcum

albomyces and Rhizomucor pusillus from Jordanian soils [44]. Several studies reported these previous fungi as

thermophilic and thermotolerant from different habitats and sources [1, 4, 5, 41, 43, 45, 51].

The water was the poorest source in fungi yielding 4 genera and 5 species and this is in agreement with the

previously reported results during 2006/2007 [9]. Thermoascus aurantiacus var. levisporus and Thermomyces

ibadanensis were isolated from the mud and soil for the first time in Egypt (Table 3.7). A. turcosus,

Chaetomium thermophile, Corynascus sepedonium, Emericella acristata, E. lata, E. quadrilineata, E. rugulosa,

E. variecolor var. astellata, and Paecilomyces variotii were isolated from the soil only while Acremonium

alabamensis, A. terreus, Paecilomyces aeruginosus, P. inflatus and P. zollerniae were recovered from the mud

only.

3.3 An Overview on the Biodiversity of Fungi

It is worth mentioning that, some fungal species were recovered from the three habitats while others were

isolated from one habitat only as follows: from water, Alternaria chlamydosporigena, A. roseoglobulosus, A.

terricola, A. unguis and many others, from mud, Acremonium alabamensis, Gymnascella hyalinospora, Isaria

felina, Paracremonium inflatum and many others, and from soil, Acremonium curvulum, Aspergillus candidus,

Botryotrichum piluliferum, Chaetomium globosum, C. nigricolor, Clonostachys rosea, C. solani and many

others.

3.4 Halophilic Ability

A total of 281 isolates related to 20 genera, 72 species and 1 variety were tested for their halophilic ability on

media supplemented with 0-30 % NaCl (Table 3.8) and these are classified as: weakly halotolerant that could

grow in 0-15 % NaCl (all isolates), moderately halotolerant in 20 % (246 isolates represented by 16 genera, 60

species and 1 variety), highly halotolerant in 25 % (32 isolates represented by 7 genera and 18 species) and

extremely halotolerant in 30 % (12 isolates represented by 2 genera and 7 species). The twelve extremely

halotolerant isolates could survive 30 % NaCl for up to 18 months, and these include Sarocladium kiliense (1

isolate AUMC 11041 from Beida mud) and the remaining 11 isolates were identified as: A. flocculosus (1

isolate AUMC 11041 from Zugm mud), A. insulicola (3 isolates; AUMC 11036 from Fasida water, AUMC

11044 from Beida soil and AUMC 11046 from Gaar soil), Aspergillus ochraceus (1 isolate; AUMC 11035 from

Fasida water), A. roseoglobulosus (2 isolates; AUMC 11037 from Umm-Rishah water and AUMC 11039 from

Hamra water), A. sclerotiorum (3 isolates; AUMC 11038 from Khadra water, AUMC 11042 from Rosetta soil

and AUMC 11045 from Beida soil), and Aspergillus species section Wentii near to A. terricola var. americana

(1 isolate; AUMC 11043 from Rosetta soil) (Table 3.8). The current results are almost in agreement with those

previously obtained results [47] where the tested 100 species and 3 varieties were categorized into 4 groups:

halophilic (grew on 5-25 % NaCl), highly halotolerant (0-20 %), moderately halotolerant (0-15 %) and weakly

halotolerant (0-10 %).

In agreement with the present results, the isolated fungal species from Cabo Rojo solar saltern were found to

grow in 0 - 25 % NaCl; however, none of them can be classified as halophilic because they all can grow in



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NaCl-free media [13]. Based on their growth rate in the presence of 25 % NaCl, they classified as highly

halotolerant species (with a growth rate >3 cm over 10 d in 25 % NaCl), moderately halotolerant species (with a

growth rate between 2 - 3 cm over 10 d in 25 % NaCl) and weakly halotolerant species (with a growth rate < 2

cm over 10 d in 25 % NaCl and can grow in up to 15 % NaCl [13].

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Table 3.1 Chemical analysis of 40 water samples collected from lakes of Wadi-El-Natrun in February 2012

(5 samples from each lake)

Lake pH % TDS Na+

K+

CO3-2

HCO3-

Ca+2

Mg+2

Cl-

CFUs G S

F 9.54±0.06 6.7±0.36 8.5±1.27 0.066±0.01 38.0±6.0 29.2±6.67 0.04±0.02 0.03±0.02 2.55±0.16 143 11 25

U 9.1±0.24 26.3±2.26 17.64±6.4 0.23±0.06 38.1±12.15 16.9±8.9 0.03±0.01 0.02±0.01 13.5±0.33 80 6 8

R 8.5±0.11 9.8±3.25 9.3±2.26 0.057±0.03 2.2±1.016 2.4±1.8 0.1±0.02 0.02±0.01 12.3±1.34 184 15 25

H 9.3±0.06 11.6±2.5 10.6±1.7 0.22±0.04 29.8±6.66 16.8±5.63 0.04±0.01 0.02±0.01 5.9±1.11 15 4 8

Z 9.27±0.02 14.4±0.8 12.5±4.45 0.45±0.6 34.2±2.066 25.8±6.26 0.1±0.01 0.03±0.01 11.3±0.34 21 3 5+1

B 8.9±0.1 4.0±0.4 3.4±0.43 0.05±0.01 1.1±0.36 2.8±0.96 0.35±0.06 0.03±0.01 1.7±0.1 63 6 11

K 9.3±0.1 1.9±0.08 1.35±0.14 0.02±0.006 4.5±2.0 5.96±1.84 0.04±0.02 0.01±0.01 0.7±0.1 56 3 8

G 8.7±0.2 1.95±0.56 1.13±0.43 0.023±0.01 0.4±0.1 1.38±0.33 0.6±0.1 0.1±0.02 0.8±0.1 30 4 9

Table 3.2 Chemical analysis of 40 mud samples collected from lakes of Wadi-El-Natrun in February 2012

(5 samples from each lake)

Lake pH % OM % MC % TDS Na+

K+

CO3-2

HCO3-

Ca+2

Mg+2

Cl-

CFUs G S

F 10.24±0.1 0.45±0.1 21.8±4.9 8.96±1.9 173.7±25.2 1.31±0.3 4.0±1.55 7.37±1.94 0.4±0.1 0.2±0.1 4.2±0.6 4062 12 20

U 10.0±0.2 4.74±0.9 22.97±2.57 6.75±1.97 121.35±59.1 6.6±4.6 0.4±0.46 3.63±4.13 0.54±0.1 0.12±0.04 27.34±8.9 5006 7 14

R 8.8±0.26 3.3±1.75 58.5±21.2 20.36±1.7 156.7±11.6 1.85±0.9 22.98±6.6 16.1±3.45 2.3±0.5 0.15±0.1 90.9±11.3 29510 10 17

H 9.74±0.3 5.7±1.7 27.3±3.99 12.6±3.12 185.7±15.2 7.1±1.9 3.9±6.4 7.7±6.25 2.2±0.7 0.15±0.1 59.3±3.98 4552 9 14

Z 9.3±0.7 5.94±2.9 48.0±14.1 12.1±2.9 182.2±35.85 1.85±0.9 5.9± 6.1 9.26±6.13 0.2±0.1 0.05±0.03 47.75±17.0 29098 12 19+1

B 9.8±0.2 0.9±0.6 24.8±4.6 18.2±6.25 281.1±139.5 3.74±1.2 12.6±8.35 10.2±3.45 0.97±0.4 0.13±0.1 59.9±16.0 7346 14 24+2

K 9.6 ± 0.2 0.5±0.5 17.56±4.3 0.66±0.4 76.5±87.9 0.97±0.3 0.12±0.1 1.67±0.75 0.28±0.1 0.1±0.04 2.5±1.4 9306 7 16+1

G 9.0±0.5 5.14±3.5 29.5±11.9 5.88±4.0 114.24±37.95 3.9±1.57 0.32±0.4 1.8±0.84 0.55±0.13 0.1±0.05 19.3±15.5 13480 9 15

Table 3.3 Chemical analysis of 33 soil samples collected from around lakes of Wadi-El-Natrun in February

2012

Lak

e(n)

pH %

OM

%

MC

%

TDS

Na+

K+

CO3-2

HCO

3-

Ca+2

Mg+2

Cl-

CF

Us

G S



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F(3) 7.8±

0.15

1.5±

0.8

12.6

±4.1

0.12

±0.0

3

13.2±

14.14

1.4±

0.9

0.02

±0.0

2

0.6±

0.06

0.6±

0.25

0.06

±0.0

1

0.8±

0.26

638

60

1

6

32

U(4

)

7.9±

0.22

1.97

±0.5

8.32

±3.2

6

0.96

±0.0

6

24.6±

6.9

2.35

±0.6

0.02

±0.0

5

0.64

±0.1

5

1.97

±2.0

1

0.04

±0.0

3

2.02

±0.5

3

513

28

1

9

30

+2

R(4

)

7.97

±0.0

5

2.06

±0.5

9.71

±4.9

3

0.33

±0.2

3

7.9±5.

1

1.4±

0.9

0.01

±0.0

2

0.55

±0.1

3

0.5±

0.26

0.04

±0.0

2

1.22

±0.7

4

670

16

1

4

21

+1

H(4

)

8.0±

0.05

2.35

±0.8

9.33

±4.8

4

0.27

±0.1

9

6.9±1.

6

0.6±

0.2

0.02

±0.0

2

0.65

±0.1

0.65

±0.5

7

0.07

±0.0

2

0.7±

0.44

457

96

1

4

24

+1

Z(3) 8.1±

0.01

2.5±

0.6

6.97

±3.2

2

0.12

±0.0

4

8.9±1.

8

0.7±

0.2

0.03

±0.0

3

0.56

±0.1

2

0.43

±0.1

5

0.06

±0.0

1

0.99

±0.7

5

642

30

1

4

23

+1

B(5

)

8.12

±0.2

5

2.3±

0.5

10.6

±3.8

0.33

±0.1

4

8.7±3.

52

1.0±

0.3

0.04

±0.0

3

0.63

±0.1

5

0.54

±0.2

0.04

±0.0

3

1.0±

0.6

658

54

1

5

27

+3

K(3

)

7.99

±0.2

5

1.9±

0.3

6.9±

2.4

0.3±

0.02

7.5±2.

23

0.84

±0.7

0.06

±0.0

3

0.53

±0.0

9

0.23

±0.0

8

0.05

±0.0

2

0.85

±0.3

4

348

00

9 21

+1

G(7

)

8.1±

0.2

2.4±

0.5

8.84

±2.9

0.6±

0.4

14.9±

7.6

1.5±

0.8

0.02

±0.0

2

0.7±

0.15

1.7±

1.56

0.05

±0.0

2

1.1±

0.8

127

390

1

3

32

+1

OM, MC and TDS are determined as %, the anions and cations are determined as g/l water and g/kg dry wt. mud

or soil; figures in Tables are mean of triplicate of number of water, mud or soil samples collected from each lake.

*G = number of genera, S = number of species and varieties, n = number of samples; F=Fasida, U=Umm-Rishah,

R=Rosetta, H=Hamra, Z=Zugm, B=Beida, K=Khadra and G=Gaar.

Table 3.4 Assiut University Mycological Centre accession number (AUMC) of fungi isolated from mud, water and

soil of different lakes of Wadi-El-Natrun with their accession GenBank numbers given together with the closest

match in the GenBank data base and sequence similarity in percent to the match as inferred from blastn searches

of ITS sequences.

Lake Isolation

medium

AUM

C

GenBank

accession

No

bp Closest match Similarity (%) Identification

Mud

Khadra Cz + 10 %

NaCl

11005 KX376276 582

KC466536

KC987176=CBS

49071T

570/580 (98.27)

494/500 (98.80)

Acremonium zonatum

Emericellopsis pallida

Zugm Cz + 10 %

NaCl

11006 KX376277 581

KC466536

KC987176=CBS

49071T

576/582 (98.96)

494/500 (98.80)

Acremonium zonatum


Beida Cz + 10 %

NaCl

11007 KX384651 582

KC466536

KC987176=CBS

49071T

573/582 (98.40)

494/500 (98.80)

Acremonium zonatum


Beida Cz + 10 %

NaCl

11008 KX376278 580

KC466536

KC987175=CBS

49171T

577/580 (99.48)

497/499(99.59)

Acremonium zonatum

Emericellopsis maritime

Fasida 1 % glucose-

Cz

11027 KX384652 615 HM991271=NRRL

2881T

590/591(99.83) Gymnascella

hyalinospora

Umm-

Rishah

1 % glucose-

Cz

11028 KX384653 627 HM991271=NRRL

2881T

590/591 (99.83) Gymnascella

hyalinospora

Rosetta 1 % glucose-

Cz

11029 KX384654 571 HG965031=MUCL

9939T

551/551(100) Sarocladium subulatum

Hamra 1 % glucose-

Cz

11030 KX384655 557 KM231829=CBS

48577T

540/557 (96.94) Paracremonium inflatum

Hamra 1 % glucose- 11031 KX384656 582 KC466536 578/581 (99.48) Acremonium zonatum



pg. 356

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Lake Isolation

medium

AUM

C

GenBank

accession

No


Cz NR_077122=CBS

63873T

563/581 (96.90) Acremonium

dichromosporum

Hamra 1 % glucose-

Cz

11032 KX384657 581

KC466536

KF032659=DM12

573/580 (98.79)

572/579 (98.79)

Acremonium zonatum


Zugm 1 % glucose-

Cz

11033 KX384658 582 NR_130684=MUCL

9724T

536/538 (99.62) Sarocladium kiliense

Zugm 1 % glucose-

Cz

11034 KX384659 562 LN864540=Kw63-15

NR_130684=MUCL

9724T

556/556 (100)

532/534 (99.62)

Sarocladium kiliense

Fasida 1 % glucose-

Cz

10329 KX531010 598 IC-A3 = HQ285562

LC105695=DY10.1.1

NR_135395=NRRL

447T

595/598(99.49)

594/597(99.49)

574/577(99.48)

A. oryzae

Water

Fasida Cz + 10 %

NaCl

11001 KX384660 581

KC466536

KF032659=DM12

KC987176=CBS

49071T

575/580 (99.10)

574/579 (99.13)

494/500 (98.80)

Acremonium zonatum


Rosetta Cz + 10 %

NaCl

11002 KX384661 473

KT968535=JCKQF3

KC987176=CBS

49071T

462/472 (97.88)

440/451(97.56)

Acremonium zonatum


Rosetta Cz + 10 %

NaCl

11003 KX384662 580 KC466536

KC987171=CBS

127350T

KC987176=CBS

49071T

570/580 (98.27)

498/499 (99.79)

494/500 (98.80)

Acremonium zonatum

Emericellopsis alkalina


Beida Cz + 10 %

NaCl

11004 KX384663 580 KC466536

KC987171=CBS

127350T

KC987176=CBS

49071T

570/580 (98.27)

498/499(99.79)

494/500(98.80)

Acremonium zonatum

Emericellopsis alkalina


Fasida 1 % glucose-

Cz

11009 KX384664 563 KJ443241=M27T 491/492 (99.79) Chordomyces

antarcticum

Fasida 1 % glucose-

Cz

11010 KX446769 557 KP068972=WM

07.196

JX508810=LC1990T

551/556 (99.10)

547/556 (98.38)

Plectosphaerella

cucumerina

Plectosphaerella

oligotrophica

Fasida 1 % glucose-

Cz

11011 556 KJ443241=M27T 487/492 (98.98) Chordomyces

antarcticum

Fasida 1 % glucose-

Cz

11012 KX385856 561 KJ443241=M27T 490/492 (99.59) Chordomyces

antarcticum

Fasida 1 % glucose-

Cz

11013 KX446768 559 FM955449=FMR

9523T

474/565 (83.89) Ramophialophora

humicola


Cz

11014 KX446755 557 KU204705=GL11101

616

JX508810=LC1990T

541/546 (99.08)

541/555 (97.47)

Plectosphaerella

cucumerina

Plectosphaerella

oligotrophica


Cz

11015 KX446756 603 KC466540

KF993392=PAV-M

1.138

591/602 (98.17)

579/588 (98.46)

Alternaria

chlamydosporigena

Alternaria

chlamydospora


Cz

11016 KX446757 581 KC466536

KC987175=CBS

49171T

KC987176=CBS

49071T

572/581 (98.45)

496/499(99.39)

492/500(98.40)

Acremonium zonatum

Emericellopsis maritime




pg. 357

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Lake Isolation

medium

AUM

C

GenBank

accession

No



Cz

11017 KX446758 581

KC466536

KC987175=CBS

49171T

KC987176=CBS

49071T

574/580 (98.96)

495/499 (99.19)

491/500 (98.20)

Acremonium zonatum

Emericellopsis maritima



Cz

11018 KX446759 582

KC466536

KC987175=CBS

49171T

KC987176=CBS

49071T

575/581 (98.96)

495/501 (98.80)

492/501(98.20)

Acremonium zonatum




Cz

11019 KX446760 580

KC466536

KC987175=CBS

49171T

KC987176=CBS

49071T

576/580 (99.31)

497/499 (99.59)

493/500 (98.60)

Acremonium zonatum




Cz

11020 KX446761 602

KC466540

KF993392=PAV-M

NR_136039=CBS

49172T

594/601 (98.83)

580/588 (98.63)

537/544 (98.71)

Alternaria

chlamydosporigena

Alternaria

chlamydospora


Cz

11021 KX446762 582

KC466536

KF032659=DM12

570/580 (98.27)

569/579 (98.27)

Acremonium zonatum



Cz

11022 KX446763 578

KC254090=UOAHCP

F 13355

KT878352=07739

565/574 (98.43)

566/576 (98.26)

Sarocladium strictum

Acremonium

sclerotigenum

Umm-

Rishah

1 % glucose-

Cz

11023 KX446764 569 FJ430712=MH178

AY952467=D99026

568/569 (99.82)

566/568 (99.64)

Stilbella fimetaria

Gaar 1 % glucose-

Cz

11024 KX446765 577 KC254091=UOAHCP

F 13842

KF225143=BAFSCH

571/575 (99.30)

571/577 (98.96)

Sarocladium strictum

Acremonium alternatum

Khadra 1 % glucose-

Cz

11025 KX446766 564 KT192193=00018-1

KT878352=07739

560/561 (99.80)

559/560 (99.82)

Acremonium alternatum

Acremonium

sclerotigenum

Beida 1 % glucose-

Cz

11026 KX446767 580 LN864540=Kw63-15

AY138846=UW 940

NR130684=MUCL

9724T

574/579 (99.13)

573/578 (99.13)

529/531 (99.62)

Sarocladium kiliense

Soil

Beida Cz+10 %

NaCl

10331 KX531011 599 AD-B3=HQ285520

CJ-B4=HQ285545

NR_111041=ATCC16

883T

599/599(100)

598/598(100)

593/596(99.49)

A. flavus

Table 3.5 Percent CFUs (calculated to the total fungi per 600 ml water or 40 g dry mud or 33 g dry soil in all

samples) and frequency (F) of fungi isolated on Cz agar at 25° C from 40 water, 40 mud and 33 soil samples

collected from the 8 lakes of Wadi-El-Natrun.

Taxa

Water Mud Soil

%CFUs F %CFUs F %CFUs F

Acremonium Link 22.84 20M 10.8 6L 0.77 2L

A. alternatum Link 0.6 4L

A. curvulum W. Gams 0.77 2L

A. zonatum (Sawada) W. Gams 7.41 3L 6.6 6L

Acremonium spp. 14.83 6L 4.2 4L

Alternaria 1.6 4L 0.3 1L 0.2 2L

A. alternata (Fries) Keissler 1.0 2L 0.3 1L 0.2 2L

A. chlamydosporigena Woudenberg & Crous 0.6 2L



pg. 358

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Taxa

Water Mud Soil


Aspergillus P. Micheli ex Link 36.5 24H 52.2 32H 56.81 33H

A. aegyptiacus Moubasher & Moustafa 0.1 1L

A. brasiliensis Varga et al. 1.2 3L 0.7 5L 6.1 13M

A. candidus Link 0.04 1L

A. deflectus Fennell & Raper 0.2 2L 0.1 1L 0.04 1L

A. flavipes (Bainier & Sartory) Thom & Church 3.2 3L 11.2 1L 0.23 3L

A. flavus Link 5.6 12L 6.67 21H

A. flavus var. columnaris Raper & Fennell 1.0 1L 0.1 1L 1.1 2L

A. foetidus (Nakaz.) Thom & Raper 0.2 1L 0.04 1L

A. fumigatus Fresenius 7.21 6L 3.7 8L 0.74 4L

A. insulicola Montemayor & Santiago 1.0 1L

A. lacticoffeatus Frisvad & Samson 0.2 1L 0.11 1L

A.neoafricanus Samson, Peterson, Frisvad & Varga 1.0 2L 12.4 1L 5.0 2L

A. niger van Tieghem 3.8 8L 5.16 14M

A. ochraceus Wilhelm 3.2 3L 1.0 5L 0.9 3L

A. oryzae (Ahlburg) E. Cohn 0.3 1L

A. parasiticus Speare 0.2 1L 4.1 8L

A. petrakii Vӧrӧs- Felkai 0.2 2L 0.31 1L

A. roseoglobulosus Frisvad & Samson 1.2 2L

A. sclerotiorum G. A. Huber 0.4 1L

A. sulphureus (Fresen.) Wehmer 6.61 2L 0.3 1L

A. sydowii (Bainier & Sartory) Thom & Church 1.8 4L 0.1 1L 0.2 3L

A. terreus Thom 6.0 4L 12.8 16M 26.15 26H

A. terricola E. J. Marchal 0.8 2L

A. unguis (Emile-Weil & Gaudin) Thom & Raper 0.8 1L

Botryotrichum piluliferum Saccardo & Marchal 1.32 2L

Chaetomium Kunze 0.23 2L

C. globosum Kunze 0.15 1L

C. nigricolor L.M. Ames 0.08 1L

Chordomyces antarcticum Bilanenko, Georgieva, Gum-

Grzhimaylo

4.21 2L

Cladosporium Link 0.4 2L 0.7 3L 0.23 3L

C. cladosporioides (Fresenius) de Vries 0.2 1L 0.2 1L 0.11 2L

C. sphaerospermum Penzig 0.2 1L 0.5 3L 0.15 1L

Cladosporium spp.

Clonostachys Corda 1.8 3L

C. rosea (Link) Schroers, Samuels, Seifert & W.

Gams

1.5 2L

C. solani (Harting) Schroers & W. Gams 0.31 1L

Cochliobolus tuberculatus Sivanesan 1.8 2L 0.6 1L 0.7 2L

Corynascus sepedonium (C.W.Emmons) Arx 0.62 1L

Emericella Berkeley & Broome 0.1 1L 16.1 22H

E. dentata (D.K.Sandhu & R.S.Sandhu) Y. Horie 3.26 3 L

E. lata Subramanian 0.3 2L

E. nidulans (Eidam) Vuillemin 0.1 1L 5.1 22H

E. quadrilineata (Thom & Raper) C.R. Benj 7.2 5L

E. rugulosa (Thom & Raper) C.R. Benj 0.31 1L

Exerohilum rostratum (Drechsler) Leonard & Suggs 0.23 1L

Fusarium Link 2.0 2L 5.9 2L 9.4 12M

F. oxysporum Schlechtendahl & Hansen 2.33 5L

F. sambucinum Füchel 1.9 1L 0.62 3L

F. semitectum Berkeley & Ravenel 1.2 2L

F. solani (Mart.) Appel & Wollenw. emend. Sny. &

Hans.

0.8 1L 4.0 1L 6.25 10L

F. thapsinum Klittich, Leslie, Nelson & Marasas 0.2 1L



pg. 359

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Taxa

Water Mud Soil


Gliocladium Corda 0.62 2L

G. catenulatum Gilman & Abbott 0.11 1L

G. penicillioides Corda 0.5 1L

Gymnascella hyalinospora (Kuehn, Orr & Ghosh)

Currah

0.3 1L

Humicola fusco-atra Traaen 0.2 1L 1.67 4L

Isaria felina (DC.) Fries 0.1 1L

Microascus manginii (Loubiѐre) Curzi 0.2 1L

Mucor Fresenius 2.0 4L 0.5 2L 0.04 1L

M. circinellioides van Tieghem 1.0 3L 0.1 1L

M. hiemalis Wehmer 1.0 2L 0.4 2L 0.04 1L

Myrothecium verrucaria (Albertini & Schweinitz)

Ditmer ex Steudel

0.15 1L

Papulaspora irregularis H.H. Hotson 0.11 1L

Paracremonium inflatum Lombard & Crous 0.3 1L

Penicillium Link 11.42 21H 15.6 19M 8.26 9L

P. aurantiogriseum Dierckx 3.0 4L 0.1 1L 3.9 6L

P. brevicompactum Dierckx 0.2 1L 0.4 1L

P. chrysogenum Thom 3.61 4L 6.8 9L 1.86 2L

P. citrinum Thom 0.3 1L

P. corylophilum Dierckx 0.04 1L

P. crustosum Thom 0.6 3L 2.4 2L 0.46 1L

P. dendriticum Pitt 0.2 1L

P. donkii Stolk 0.2 1L

P. duclauxii Delacroix 0.1 1L 0.43 2L

P. funiculosum Thom 1.6 3L

P. griseofulvum Dierckx 0.2 1L

P. italicum Wehmer 0.6 1L

P. janthinellum Biourge 1.0 3L

P. melinii Thom 0.6 2L

P. olsonii Bainier & Sartory 1.0 1L

P. oxalicum Currie & Thom 2.7 2L

P. puberulum Bainier 0.3 2L

P. raistrickii G.Sm. 0.1 1L

P. sublateritium Biourge 0.2 1L

P. waksmanii K.M. Zalessky 0.6 1L

Penicillium spp. 1.8 4L

Phaeoacremonium sp. 1.2 1L

Phialophora sp. 0.2 1L 0.2 1L

Phoma glomerata (Corda) Wollenw. & Hochapfel 0.11 1L

Plectosphaerella oligotrophica Liu, Hu, Liu & Cai 2.0 1L

Purpureocillium lilacinum (Thom) Luangsa-ard,

Houbraken, Hywel-Jones & Samson

0.1 1L 0.04 1L

Ramophialophora sp. AUMC 11013 0.2 1L

Sarocladium Gams & Hawksworth 4.41 6L 5.6 3L

S. kiliense (Grütz) Summerbell 0.2 1L 3.4 2L

S. strictum (W. Gams) Summerbell 4.21 5L 1.6 1L

S. subulatum Giraldo, Gené & Guarro 0.6 1L

Scopulariopsis Bainier 5.0 7L 1.4 3L

S. acremonium (Delacroix) Vuillemin 0.2 1L

S. brevicaulis (Saccardo) Bainier 1.6 3L 0.1 1L

S. brumptii Salv. -Duval 1.0 1L 0.1 1L

S. carbonaria Morton & G. Sm. 1.8 2L

S. fusca Zach 0.4 1L 1.2 1L

Stachybotrys Corda 0.4 4L



pg. 360

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Taxa

Water Mud Soil


S. dichroa Grove 0.1 1L

S. ramosa Udaiyan 0.1 1L

S. verrucispora Matsushima 0.1 1L

S. virgata Krzemien. & Badura 0.1 1L

Stilbella fimetaria (Pers.) Lindau 0.2 2L

Trichoderma Pers. 3.2 4L 4.9 8L 0.15 2L

T. harzianum Rifai 2.0 3L 4.6 7L 0.15 2L

T. koningii Oudem 1.0 1L

T. longibrachiatum Rifai 0.2 2L 0.3 3L

Ulocladium Preuss 0.4 2L 0.43 2L

U. atrum Preuss 0.43 2L

U. botrytis Preuss 0.2 1L

U. tuberculatum E.G. Simmons 0.2 1L

Total CFUs (615899) 20 28 22

No. of genera 35 & species 105 + 1 variety 61 + 1 variety 50 + 1 variety 53 + 1 variety

Table 3.6 Percent CFUs (calculated to the total fungi per 600 ml water or 40 g dry mud or 33 g dry soil in

all samples) and frequency (F) of fungi isolated on Cz + 10 % NaCl at 25° C from 40 water, 40 mud and 33

soil samples collected from the 8 lakes of Wadi-El-Natrun.

Taxa Water Mud Soil


Acremonium Link 93.48 8L 6.7 7L

A. zonatum 10.87 2L 3.16 7L

Acremonium spp. 82.61 7L 3.56 5L

Aspergillus 85.46 13M 81.0 31H

A. aegyptiacus 0.1 1L 0.12 1L

A. brasiliensis 0.1 1L

A. flavipes 36.8 3L 0.12 1L

A. flavus 18.9 9L 37.2 9L

A. flavus var. columnaris 2.99 4L

A. flocculosus Frisvad & Samson 7.7 1L

A. fumigatus 0.1 1L

A. insulicola 4.5 3L

A. neoafricanus 1.5 1L

A. niger 0.25 1L

A. ochraceus 0.75 5L

A. parasiticus 3.74 7L

A. petrakii 3.24 2L

A. sclerotiorum 1.37 3L

A. sydowii 3.76 4L

A. tamarii Kita 0.1 1L 2.62 7L

A. terreus 0.12 1L

A. versicolor (Vuillemin) Tirab. 18.0 9L 22.22 13M

Aspergillus sp. near to A. terricola var. americana 0.12 1L

Cladosporium sp. 0.12 1L

Emericella 0.5 1L 2.62 5L

E. nidulans 0.5 1L 0.37 3L

E. quadrilineata 2.25 4L

Eurotium Link 0.75 2L

E. amstelodami (Mangin) Thom & Church 0.25 1L

E. rubrum Thom & Church 0.5 1L

Fusarium 0.4 2L 5.61 3L

F. semitectum 5.37 2L

F. solani 0.4 2L 0.25 1L

Microascus manginii 0.25 2L

Penicillium 2.17 1L 1.4 4L 9.36 14M



pg. 361

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Taxa Water Mud Soil


P. aurantiogriseum 0.25 2L

P. chrysogenum 2.17 1L 1.3 3L 7.24 12M

P. donkii 0.1 1L

P. funiculosum 1.87 5L

Sarocladium 4.94 6L

S. kiliense 4.55 5L

S. strictum 0.4 2L

Scopulariopsis 4.35 2L 0.37 2L

S. brevicaulis 0.12 1L

S. brumptii 0.25 1L

S. halophilica Tubaki 4.35 2L

Ulocladium lanuginosum (Harz.) Simmons 0.4 1L

Total CFUs (3670) 100 (46) 100 (2022) 100 (1602)

No. of genera 11 & species 38 + 1 variety 3 & 3 8 & 17 7 & 27 + 1

Table 3.7 Percent CFUs (calculated to the total fungi per 600 ml water or 40 g dry mud or 33 g dry soil in

all samples) and frequency (F) of fungi isolated on YpSs at 45° C from 40 water, 40 mud and 33 soil samples

collected from the 8 lakes of Wadi-El-Natrun.

Taxa Water Mud Soil


Acremonium 2.36 4L 0.06 1L

A. alabamensis Morgan-Jones 0.6 1L

A. thermophilum W. Gams & J. Lacey 1.2 3L 0.06 1L

Acremonium sp. (chlamydospores) 0.6 1L

Aspergillus 78.72 15M 30.2 15M 17.76 25H

A. fumigatus 76.6 13M 28.4 13M 17.3 23H

A. terreus 0.6 1L

A. turcosus Hong, Frisvad & Samson 0.26 1L

A. viridinutans Ducker & Thrower 2.13 1L 1.2 1L 0.2 1L

Chaetomium thermophile La Touche 2.53 1L

Corynascus sepedonium (C.W.Emmons) Arx 0.77 1L

Emericella Berkeley & Broome 0.6 1L 25.92 9L

E. acristata Fennell & Raper 0.06 1L

E. lata 1.68 1L

E. nidulans 0.6 1L 8.88 2L

E. quadrilineata 14.45 6L

E. rugulosa 0.77 1L

E. variecolor var. astellata (Fennell & Raper)

Benjamin

0.06 1L

Humicola grisea var. thermoidea Cooney & Emerson 1.2 3L 2.33 1L

Malbranchea cinnamomea (Lib.) Oorschot & de Hoog 2.13 1L 8.9 7L 23.4 21H

Myriococcum albomyces Cooney & Emerson 0.6 1L 3.7 7L

Paecilomyces Bainier 3.55 4L 0.6 2L

P. aeruginosus 0.6 1L

P. inflatus (Burnside) J.W. Carmich. 0.6 1L

P. variotii Bainier 0.6 2L

P. zollerniae Stolk & Samson 0.6 1L

Paecilomyces spp. 1.77 2L

Rhizomucor pusillus (Lindt) Schipper 2.36 2L 1.0 7L

Scytalidium thermophilum (Cooney & Emerson)

Austwick

11.24 4L 5.38 2L

Talaromyces thermophilus Stolk 4.25 1L 21.9 6L 4.3 8L

Thermoascus aurantiacus var. levisporus Upadhyay,

Farmelo, Goetz & Melan

1.18 2L 0.13 1L

Thermomyces Tsikl 14.9 2L 15.97 8L 12.12 20M

T. ibadanensis Apinis & Eggins 4.14 1L 4.5 7L



pg. 362

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Taxa Water Mud Soil


T. lanuginosus Tsikl 14.9 2L 11.83 8L 7.65 18M

Total CFUs (3471) 100 (47) 100 (338) 100 (3086)

No. of genera 14 & species 25 + 3 varieties 4 & 5 12 & 18 + 2 14 & 19 + 3

Table 3.8 Number of isolates of fungal genera and species tested for their halophilic abilities on Cz

medium supplemented with different NaCl concentrations

Taxa isolates tested 0-15% 20 % 25 % 30 %

Acremonium 18 18 16 3

A. alternatum 1 1 1

A. zonatum 17 17 15 3

Alternaria 3 3 2

A. alternata 1 1

A. chlamydosporigena 2 2 2

Aspergillus 139 139 132 21 11

A. aegyptiacus 1 1 1

A. brasiliensis 5 5 4

A. deflectus 1 1

A. flavipes 12 12 12

A. flavus 30 30 30 3

A. flavus var. columnaris 6 6 6

A. flocculosus 1 1 1 1 1

A. foetidus 1 1 1

A. fumigatus 8 8 4

A. insulicola 3 3 3 3 3

A. lacticoffeatus 1 1 1

A.neoafricanus 2 2 2

A. niger 2 2 1

A. ochraceus 9 9 9 1 1

A. parasiticus 7 7 7

A. petrakii 2 2 2

A. roseoglobulosus 2 2 2 2 2

A. sclerotiorum 3 3 3 3 3

A. sulphureus 4 4 4

A. sydowii 12 12 12 4

A. tamarii 1 1 1

A. terreus 22 22 22 2

A. terricola 1 1 1 1

A. unguis 1 1 1

A. versicolor 1 1 1

Aspergillus sp. near to A. terricola var. americana 1 1 1 1 1

Chordomyces antarcticum 2 2 2

Cladosporium 2 2

C. cladosporioides 1 1

C. sphaerospermum 1 1

Cochliobolus tuberculatus 4 4 4

Emericella 8 8 8

E. nidulans 4 4 4

E. quadrilineata 4 4 4

Eurotium 5 5 5 1

E. amstelodami 2 2 2

E. rubrum 3 3 3 1

Fusarium 13 13 13 1

F. semitectum 8 8 8 1

F. solani 5 5 5

Humicola fusco-atra 1 1 1

Microascus manginii 2 2 2



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Taxa isolates tested 0-15% 20 % 25 % 30 %

Mucor 4 4

M. circinellioides 3 3

M. hiemalis 1 1

Penicillium 50 50 48 3

P. aurantiogriseum 7 7 7

P. brevicompactum 1 1 1

P. chrysogenum 24 24 24 1

P. crustosum 3 3 3 1

P. dendriticum 1 1 1

P. donkii 2 2 1

P. funiculosum 5 5 5

P. griseofulvum 1 1 1

P. italicum 1 1 1

P. melinii 2 2 2

P. olsonii 2 2 1

P. sublateritium 1 1 1 1

Plectosphaerella oligotrophica 2 2 2

Ramophialophora near to humicola 1 1 1

Sarocladium 6 6 4 3 1

S. kiliense 2 2 1 1 1

S. strictum 4 4 3 2

Scopulariopsis 10 10 7

S. acremonium 1 1 1

S. brevicaulis 2 2 1

S. brumptii 4 4 3

S. carbonaria 1 1 1

S. fusca 1 1

S. halophilica 1 1 1

Stilbella fimetaria 1 1

Trichoderma 7 7

T. harzianum 2 2

T. koningii 4 4

T. longibrachiatum 1 1

Ulocladium 3 3 1

U. botrytis 1 1

U. lanuginosum 1 1

U. tuberculatum 1 1 1

Total 281 281 248 32 12

No. of genera (20) 20 16 7 2

No. of species (72 + 1 variety) 72+1 variety 60+1 18 7

issn no.: 2454- 2024 (online) international journal of ... fungi and... · widely [57]. dead sea,...

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